Animal-type sphingoglycolipids are utilized as chemical products in raw materials of medicines and cosmetics. Sugar chains serve as receptors for various viruses including influenza virus and bacterial toxins (Non-Patent Document 1), and are therefore prospective as a novel remedy for these infectious diseases in early-stage practical application, which does not depend on antibiotics and chemically synthesized medical materials (Patent Documents 1 and 2). For example, it is known that ceramide trihexoside is a receptor for Vero toxin (toxin produced by Escherichia coli O-157) and Shiga toxin (toxin produce by dysentery bacilli) to bind to cell surface (Non-Patent Documents 2 and 3), and that lactosylceramide is a receptor for gonococci and Propionibacterium (bacteria causing dermatitis) to infect cells (Non-Patent Documents 4 and 5). However, conventional production methods which collect materials from animal brain have a problem of infectious diseases such as BSE (bovine spongiform encephalopathy), and artificial synthesis has the drawback of being difficult and costly.
Examples of research on the production of useful substances by using genetically modified plants have been recently reported (Non-Patent Documents 6 to 10). The merits of those methods are low cost, absence of carbon dioxide release and no risk of contamination by animal infectious diseases.
The fundamental structure of almost all species of sphingoglycolipids is lactosylceramide, and the variety of sphingoglycolipids, which is said to have more than 300 species, are biosynthesized in animal tissues by adding sugar chains to lactosylceramide. It is known that lactosylceramide can be synthesized from its precursor, glucosylceramide, by a sugar transfer reaction with β1,4-galactosyltransferase (β1,4GT) (Patent Document 3). Although plants have glucosylceramide, they cannot produce lactosylceramide due to the lack of β1,4GT, and as a result, they cannot produce animal-type glycolipids.
A research example reported the transfer of galactose to proteins by introducing hβ-1,4-GalT1 (Accession Number: X55415 or X13223), which is an isozyme of β1,4GT, into tobacco cells (not a plant body) cultured in a liquid medium (Non-Patent Document 11), but no example reports the transfer of sugars to lipids.
Technical literatures relating to the invention of the present application are shown below.    [Patent Document 1] Japanese Patent Kohyo* Publication No. 2003-535965 (*unexamined Japanese national phase publication corresponding to a non-Japanese international publication)    [Patent Document 2] Japanese Patent Kohyo Publication No. Hei 10-50347    [Patent Document 3] Japanese Patent Kokai** No. Hei 10-295371 (**unexamined, published Japanese patent application)    [Non-Patent Document 1] Karlson, K. A. Animal glycosphingolipids as membrane attachment site for bacteria. Ann. Rev. Biochem. 58, 309-350, 1989    [Non-Patent Document 2] Cohen A, Hannigan G E, Williams B R, Lingwood C A. J Biol Chem. 1987 Dec 15;262(35):17088-91. Roles of globotriosyl- and galabiosylceramide in verotoxin binding and high affinity interferon receptor.    [Non-Patent Document 3] Lindberg A A, Brown J E, Stromberg N, Westling-Ryd M, Schultz J E, Karlsson K A. J Biol Chem. 1987 Feb 5;262(4):1779-85. Identification of the carbohydrate receptor for Shiga toxin produced by Shigella dysenteriae type 1.    [Non-Patent Document 4] Stromberg N, Deal C, Nyberg G, Normark S, So M, Karlsson K A. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4902-6. Identification of carbohydrate structures that are possible receptors for Neisseria gonorrhoeae.     [Non-Patent Document 5] Stromberg N, Ryd M, Lindberg A A, Karlsson K A. FEBS Lett. 1988 May 9;232(1):193-8. Studies on the binding of bacteria to glycolipids. Two species of Propionibacterium apparently recognize separate epitopes on lactose of lactosylceramide.    [Non-Patent Document 6] Voelker, T. A. et al.: Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants, Science, 257, 72-74(1992)    [Non-Patent Document 7] Sayanova, O. et al.: Expression of a borage desaturase eDNA containing and N-terminal cytochrome b5 domain results in the accumulation of high levels of Δ6-desaturated fatty acids in transgenic tobacco, Proc. Natl. Acad. Sci. U.S.A., 94, 4211-4216(1997)    [Non-Patent Document 8] Ye, X. et al.: Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm, Science, 287, 303-305(2000)    [Non-Patent Document 9] Datta, K.: Bioengineered ‘golden’ indica rice cultivars with β-carotene metabolism in the endosperm with hygromycin and mannose selection systems, Plant Biotech. J., 1, 81-90(2003)    [Non-Patent Document 10] Tozawa, Y et al.: Characterization of rice anthranilate synthase a-subunit genes OASA1 and OASA2. Tryptophan accumulation in transgenic rice expressing a feedback-insensitive mutant of OASA1, Plant Physiology, 126, 1493-1506(2001)    [Non-Patent Document 11] Nirianne Q. Palacpac et al., Stable expression of human β1,4-galactosyltransferase in plant cells modifies N-linked glycosylation patterns, Proc. Natl. Acad. Sci. USA, 96, 4692-4697 (1999)    [Non-Patent Document 12] Watarai, S. et al., Inhibition of vero cell cytotoxic activity in Escherichia coli O157:H7 lysates by globotriaosylceramide, Gb3, from bovine milk. Biosci. Biotechnol. Biochem. 65, 414-419 (2001).    [Non-Patent Document 13] Hasegawa, A., Morita, M., Kojima, Y., Ishida, H. & Kiso, M., Synthesis of cerebroside, lactosylceramide, and ganglioside GM3 analogs containing β-thioglycosidically linked. Carbonhydr. Res. 214, 43-53 (1991).    [Non-Patent Document 14] N. Strömberg et al. (1988) Two species of Propionibacterium apparently recognize separate epitopes on lactose of lactosylceramide FEBS Lett. 232, 193-198.    [Non-Patent Document 15] K. Furukawa and T. Sato (1999) β-1,4-Galactosylation of N-glycans is a complex process. Biochim. Biophys. Acta. 1473, 54-66.    [Non-Patent Document 16] M. Amado et al. (1999) Identification and characterization of large galactosyltransferase gene families: galactosyltransferase for all functions. Biochi. Biophys. Acta. 1473, 35-53.    [Non-Patent Document 17] Harwood, J. L. (1998) What's so special about plant lipids? In Plant Lipid Biosynthesis, ed. Harwood, J. L., 1-26. Cambridge University Press.    [Non-Patent Document 18] Shimojima, M., Ohta, H., Iwamatsu, A., Masuda, T., Shioi, Y., and Takamiya, K. (1997) Cloning of the gene for monogalactosyldiacylglycerol synthase and its evolutionary origin. Proc. Natl. Acad. Asi. USA. 94, 333-337.    [Non-Patent Document 19] Miego, C., Marechal, E., Shimojima, M., Block, M. A., Ohta, H., Takamiya, K., Douce, R., and Joyard, J. (1999) Biochemichal and topological properties of type A MGDG synthase, a spinach chloroplast envelope enzyme catalyzing the synthesis of both prokaryotic and eukaryotic MGDG Eur. J. Biochem. 365, 990-1001.    [Non-Patent Document 20] Kelly, A. A., Froehlich, J. E., and Dörmann, P. (2003) Disruption of the two digalactosyldiacylglycerol synthase genes DGD1 and DGD2 in Arabidopsis reveals the existence of and additional enzyme of galactolipid synthesis. Plant Cell, 15, 2694-2706.    [Non-Patent Document 21] Bligh, E. G. and Dyer, W. J. (1959) A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37, 911-917.    [Non-Patent Document 22] Francois, C., Marshall, R. D., and Neuberger, A. (1962) Carbonhydrates in protein. 4. The determination of mannose in hen's-egg albumin by radioisotope dilution. Biochem. J., 83, 335-341.    [Non-Patent Document 23] Ohnishi, M., Ito, S., and Fujino, Y. (1983) Characterization of sphingolipids in spinach leaves. Biochim. Biophys. Acta., 752, 416-422.    [Non-Patent Document 24] Lynch, D. V. (1993) in Lipid metabolism in plants. Sphingolipids. CRC press, 285-308.    [Non-Patent Document 25] Kelly, A. A., and Dörmann, P. (2004) Green light for galactolipid trafficking. Curr. Opinions Plant Biol., 7, 262-269.    [Non-Patent Document 26] Imai, H., Ohnishi, M., Kinoshita, M., Kojima, M., and Ito, S. (1995) Structure and distribution of cerebroside containing unsaturated hydroxyl fatty acids in plant leaves. Biosci. Biotech. Biochem., 59, 1309-1313.    [Non-Patent Document 27] Sanders, P. R., Winter, J. A., Barnason, A. R., Rogers, S. G., and Fraley, R. T. (1984) Comparison of cauliflower mosaic virus 35S and nopaline synthase promoters in transgenic plants. Nucleic Acid Res., 15, 1543-1558.    [Non-Patent Document 28] Trinchera, M., Fiorilli, A., and Ghidoni, R. (1991) Localyzation in the golgi apparatus of rat liver UDP-Gal:glucosylceramide β1,4-galactosyltransferase. Biochemistry, 30, 2719-2724.